Techne PrimeQ - Manual (Page 140)

140
Bin threshold
The user can specify a temperature range (ºC) either side of a cursor in which a peak will be
considered valid. For example, if the bin threshold is set to 80°C with a range of 0.5°C, then all
peaks with a Tm between 79.5 and 80.5°C will be considered in the results. The default is 0.5°C.
Peaks within the valid +/- temperature range are sent to the peak column results table. If a peak is
present but falls outside of the valid range, then the box is left blank such that there will be as
many peak columns as there are cursors defined. Replicates are not averaged.
• If the cursors are very close together then it is possible that their validity ranges may
overlap. In this case, the space between should be shared equally so overlap no longer
occurs e.g. if there is a cursor of 80°C and one at 81°C and the validity range is 1.5°C, then
cursor 1 should have a validity range of 79.0 to 80.5°C and cursor 2, a range of 80.5 to
82.0°C.
• When the experiment is first run, there is no data shown in the results table as peaks have
yet to be defined. Once the positions of the cursor and their names have been set, these
can be saved as part of the Experiment file to allow a repeat run with the same parameters.
3.15.2.5 Peak area
When peak detection parameters have been set in the Dissociation Curve Wizard, clicking Next
leads through to the Peak Area screen.
The peak area option calculates the area under the selected peak(s) of interest and can provide
an indication of the relative amounts of each product in a sample.
The peak area of an unknown sample can be compared against the peak area of a known
standard amplified in the same tube. A Gaussian fit is performed for each peak of interest and the
area calculated is presented in a table similar to that for Tm results.